Radiation-Induced Protein Succinylation in Wild-Type and SIRT5-Deficient Murine Hearts: A Comparative Proteomics Study
Ontology highlight
ABSTRACT:
ORGANISM(S): Mus Musculus
SUBMITTER: 
Anwen Liu
PROVIDER: PXD070666 | iProX | Wed Nov 12 00:00:00 GMT 2025
REPOSITORIES: iProX
Similar Datasets
Project description:Objectives: To explore how modulation of cardiomyocyte SIRT5 levels affects the development of pathological cardiac remodeling and dysfunction. Background: SIRT5 is a mitochondrial NAD+-dependent deacylase, which regulates metabolic enzymes upon desuccinylation, demalonylation, and deglutarylation. Its role in the development of heart failure is not completely understood. Methods: Mice with cardiomyocyte-specific Sirt5 deletion (Sirt5-/-) or Sirt5 overexpression (cSirt5-Tg) underwent transverse aortic constriction (TAC) or Sham surgery. Cardiac structure and function were assessed by echocardiography and quantification of myocardial fibrosis and cardiomyocyte cell size. Metabolomic analysis and RNA sequencing were performed to identify potential novel pathways regulated by SIRT5. Results: While cardiac structure and function were similar in cSirt5-Tg compared to control mice, cSirt5-/- mice displayed exacerbated cardiac dilation, dysfunction, and fibrosis following TAC. Accumulation of metabolites involved in purine metabolism, particularly inosine and hypoxanthine, and reductions in nucleosides, adenosine and adenine, and nucleotides, AMP and ADP, were observed in hearts of cSirt5-/- mice and following TAC. Moreover, transcriptional analysis revealed upregulation of purine-nucleoside phosphorylase (PNP) and 5' nucleotidase, ecto (NT5E) and downregulation of adenosine kinase (ADK), which is likely to contribute to enhanced degradation of purine nucleotides and resulting accumulation of inosine and hypoxanthine in hearts of cSirt5-/- mice following TAC. Lastly, reduced SIRT5 expression and protein succinylation in left-ventricular tissue of patients with heart failure correlates with reduced ADK expression. Conclusions: Loss of SIRT5 aggravates cardiac remodeling and dysfunction in response to chronic pressure-overload possibly by dysregulation of genes involved in purine and pyrimidine metabolism, resulting in impaired ATP regeneration from adenosine.
2025-12-02 | GSE280863 | GEO
Project description:SIRT5 is one of the seven members of the NAD+-dependent sirtuin family of protein deacylase that is mainly present in mitochondria and regulates metabolism. In heart, SIRT5 is highly expressed and responsible for succinylation on metabolic enzymes. Although the role of SIRT5 in maintaining cardiac homeostasis under physiological stress and few potential substrates of SIRT5 have been preliminarily revealed, the regulatory network and key cellular signaling pathways involving SIRT5 in myocardial hypertrophy remain largely unknown. Here, we used an established murine model of pressure overload-induced myocardial hypertrophy caused by transverse aortic constriction (TAC) to outline the network and pathway involving SIRT5 in cardiac stress responses. Remarkably, SIRT5 KO mice had enhanced myocardial hypertrophy after TAC surgery compared with wild-type mice.
2024-10-05 | GSE186985 | GEO
Project description:We are studying the role of human sirtuin SIRT5 during viral infection with SARS-CoV-2. We performed RNA-sequencing of WT and SIRT5-KO human lung adenocarinoma A549 cells overexpressing ACE2 (A549-ACE2), in infected and mock-infected conditions. . Our analysis revealed that SARS-CoV-2 replication is attenuated in SIRT5-KO cells. In addition, SIRT5-KO cells expressed higher basal levels of innate immunity markers and mounted a stronger antiviral response. Our results indicate that SIRT5 is a proviral factor necessary for efficient viral replication.
2021-11-10 | GSE188382 | GEO
Project description:: Prostate cancer (PCa) is the most commonly diagnosed genital cancer in men worldwide. Among patients who developed advanced PCa, 80% suffered from bone metastasis, with a sharp drop in the survival rate. Using quantitative global succinylome analysis, we characterized a significant increase of lysine 118 succinylation (K118su) of lactate dehydrogenase A (LDHA), which plays a role in increasing LDHA activity. As a substrate of SIRT5, LDHA-K118su significantly increased the migration and invasion of PCa cells and LDHA activity in PCa patients.
2023-06-21 | PXD022005 | Pride
Project description:Cutaneous melanoma remains the most lethal skin cancer, and ranks third among all malignancies in terms of years of life lost. Despite the advent of immune checkpoint and targeted therapies, only roughly half of patients with advanced melanoma achieve a durable remission. SIRT5 is a member of the sirtuin family of protein deacylases that regulate metabolism and other biological processes. Germline Sirt5 deficiency is associated with mild phenotypes in mice. Here we show that SIRT5 is required for proliferation and survival across all cutaneous melanoma genotypes tested, as well as uveal melanoma, a genetically distinct melanoma subtype that arises in the eye, and is incurable once metastatic. Likewise, SIRT5 is required for efficient tumor formation by melanoma xenografts, and in an autochthonous mouse Braf;Pten-driven melanoma model. Via metabolite and transcriptomic analyses, we find that SIRT5 is required to maintain histone acetylation and methylation levels in melanoma cells, thereby promoting proper gene expression. SIRT5-dependent genes notably include MITF, a key lineage-specific survival oncogene in melanoma, and the c-MYC proto-oncogene. SIRT5 may represent a novel, druggable genotype-independent addiction in melanoma.
2021-05-07 | GSE169205 | GEO
Project description:Hepatocellular carcinoma (HCC) is a prototypical inflammation-associated cancer and the tumor microenvironment (TME) plays a pivotal role in HCC pathogenesis and response to therapy. The liver is a metabolically active organ, but how liver metabolism impacts TME during HCC development and response to immunotherapy are poorly understood. Here, we show that the metabolic regulator SIRT5 is downregulated in human primary HCC samples, and that Sirt5 deficiency in mice synergizes with oncogenes to increase bile acid (BA) production, which is due to hypersuccinylation and increased BA biosynthesis in the peroxisome. BA acts as a cell-signaling mediator to stimulate its nuclear receptor and promotes M2-like macrophage polarization, thereby creating an immunosuppressive microenvironment favorable for HCC initiating cells. Furthermore, high serum taurocholic acid, a major primary BA, correlates with low SIRT5 expression and increased M2-like tumor-associated macrophages (TAMs) in liver tissue samples from HCC patients. Importantly, administration of cholestyramine, a BA sequestrant and FDA-approved medication for hyperlipemia, reverses the effect of Sirt5 deficiency on promoting M2-like polarized TAMs and liver tumor growth. Our study thus uncovers a novel function of SIRT5 in orchestrating BA metabolism to prevent tumor immune evasion and suppress HCC. These results also suggest a potential strategy using clinically proven bile acid sequestrants for the treatment of HCC patients, especially those with decreased SIRT5 and abnormally high BAs.
2022-01-10 | GSE193213 | GEO
Project description:Chondrocyte metabolic dysfunction plays an important role in osteoarthritis (OA) development during aging and obesity. However, it is unknown how metabolic dysfunction occurs during these two conditions. Proteins post-translational modifications (PTMs) have recently emerged as an important regulator of cellular metabolism. We previously reported that one type of PTM, lysine malonylation (MaK) was increased in cartilage during obesity. Sirt5, the only known mammalian demalonylase, declines in cartilage during aging. To elucidate the interplay between Sirt5 deficiency and obesity in joint degeneration, we deploy systemic and cartilage-specific conditional knockout mouse models. Our findings reveal that the combination of Sirt5 deficiency and obesity exacerbates joint degeneration in mice. Through comprehensive proteomics analysis, we delineate the malonylome in chondrocytes, pinpointing MaK's predominant impact on various metabolic pathways such as carbon metabolism and glycolysis. Further biochemical investigation focuses on glyceraldehyde 3-phosphate dehydrogenase (GAPDH), a target of MaK, confirming a reduction in its enzymatic activity and functionality by MaK. Lastly, we identified a rare coding mutation in SIRT5 that dominantly segregates in a family with hand OA. The mutation results in substitution of an evolutionally invariant phenylalanine (Phe–F) to leucine (Leu–L) (F101L), which is located in the catalytic domain. F101L mutant results in higher MaK level and decreased expression of cartilage ECM genes (e.g., Acan and Col2a1) and upregulation of inflammation associated genes (e.g., Il6 and Nos2). In conclusion, we found that Sirt5 mediated MaK is an important regulator of chondrocyte cellular metabolism and dysregulation of Sirt5-MaK could be an important mechanism underlying aging and obesity associated OA development.
2025-12-01 | GSE272422 | GEO
Project description:A wide range of protein acyl modifications has been identified on enzymes across various metabolic processes; however, the impact of these modifications remains poorly understood. Protein glutarylation is a recently identified modification that can be non-enzymatically driven by glutaryl-CoA. In mammalian systems, this unique metabolite is only produced in the lysine and tryptophan oxidative pathways. To better understand the biology of protein glutarylation, we studied the relationship between enzymes within the lysine/tryptophan catabolic pathways, protein glutarylation, and regulation by the deglutarylating enzyme Sirtuin 5 (SIRT5). Here, we identify glutarylation of the lysine oxidation pathway enzyme glutaryl-CoA dehydrogenase (GCDH). We show increased GCDH glutarylation when glutaryl-CoA production is stimulated by lysine catabolism. Our data reveal glutarylation of GCDH impacts its function, ultimately decreasing lysine oxidation. We then demonstrate the ability of SIRT5 to deglutarylate GCDH, restoring its enzymatic activity. Finally, metabolomic and bioinformatic data indicate a novel role for SIRT5 in regulation of amino acid metabolism. Together, these data suggest a model whereby a feedback loop exists within the lysine/tryptophan oxidation pathway, in which glutaryl-CoA is produced, in turn inhibiting GCDH function. This inhibition is relieved by SIRT5 deacylation activity.
2022-02-21 | PXD018156 | Pride
Project description:Plant-pathogenic fungi have developed kinds of detoxification strategies to suppress host reactive oxygen species (ROS) for colonization, but how this process is elaborately regulated has not been well revealed. Here we show that the SIRT5-mediated protein desuccinylation acts as a master regulator to regulate ROS detoxification in the rice blast fungus Magnaporthe oryzae. The sirtuin protein SIRT5 was identified as a desuccinylase in M. oryzae, deletion of which resulted in increased sensitivity to oxidative stress. Further analysis found that Sirt5 is important for fungal virulence and adaptation to host oxidative stress. Quantitative proteomics analysis under oxidative stress identified a large number of SIRT5-mediated succinylated proteins, most of which were mitochondrial proteins involved in oxidative phosphorylation, TCA cycle, fatty acid oxidation, etc. Many succinylated proteins were enzymes involved in different ROS de-toxification processes. Disruption of SIRT5 resulted in hypersuccinylation of detoxification-related enzymes, and significant reduction of NADPH/NADP+ and GSH/GSSG ratios, disrupting redox balance and impeding the expansion of invasive hyphae in the host. Interestingly, we proved that SIRT5 can desuccinylate thioredoxin Trx2 and glutathione peroxidase Hyr1 to activate its enzyme activity, probably by affecting its proper folding. Further analysis indicated that the succinylation sites of Trx2 (K144) and Hyr1 (K101, K127) were important for their function in ROS detoxification and virulence. Altogether, this work demonstrates a novel regulatory mechanism of SIRT5-mediated desuccinylation in detoxifying host ROS during M. oryzae infection.
2024-03-06 | PXD044696 | Pride
Project description:Role of SIRT5 in Inhibition of Myocardial Hypertrophy
| PRJNA777232 | ENA